Development of a novel production platform for complex recombinant biotherapeutic

描述（由申请人提供）：基于重组蛋白的生物药物的生产是一项复杂的劳动力和资本密集型奋进。目前公认的是，使用哺乳动物细胞是生产大多数人蛋白质的必要条件，所述人蛋白质通常含有原核生物和单细胞真核生物不进行的广泛的翻译后修饰。虽然哺乳动物细胞，如中国仓鼠卵巢细胞和幼仓鼠肾细胞可以忠实地生物合成大多数人类蛋白质，但效率明显低于细菌或酵母细胞。在目前市售的重组蛋白质中，凝血因子VIII的生产效率最低，并且在单位质量基础上是迄今为止最昂贵的。其他昂贵且难以制造的生物药物包括其他凝血因子（例如因子IX和VIIa）、干扰素和基于细胞因子的细胞因子。本申请的目的是开发一种技术，该技术可以提高这些难以制造的重组生物制剂的产量和制药价格。Expression Therapeutics开发了一种新的幼仓鼠肾衍生（BHK）细胞系，命名为BHK-MS，可在悬浮生物反应器条件下在无血清和血液制品培养基中无限培养和扩增。在目前的研究中，我们1）开发和验证悬浮适应的BHK-MS细胞系，用于生产上级凝血因子VIII和因子IX，2）进行中试规模生产运行，并分析BHK-MS生物合成的重组因子VIII和因子IX的生化和药物特征，其将作为难以生产生物药物的模型。 公共卫生相关性：目前项目的长期商业目标是将基于悬浮细胞的生物制造平台商业化，该平台旨在提高复杂、难以制造的生物制品的体积生产率。目前的低产量制造工艺往往导致有限的产品可用性和高产品成本。Expression Therapeutics开发了一种正在申请专利的细胞系，旨在有效生产重组生物制剂。本文提出的研究旨在提供效率和可扩展性的概念验证，以及这种新型制造平台的严格临床前验证和表征。

DESCRIPTION (provided by applicant): The manufacturing of recombinant protein-based biopharmaceuticals is a complex, labor and capital intensive endeavor. It currently is accepted that the use of mammalian cells is a requirement for the production of most human proteins, which typically contain extensive post-translational modifications that are not performed by prokaryotes and single-celled eukaryotes. Although mammalian cells such as Chinese hamster ovary cells and baby hamster kidney cells can faithfully biosynthesize most human proteins, the efficiency is dramatically lower than is achieved by bacterial or yeast cells. Of the recombinant proteins currently marketed, coagulation factor VIII is manufactured with the lowest efficiency and is by far the most expensive on a per unit mass basis. Other expensive and difficult to manufacture biopharmaceuticals include the other coagulation factors (e.g. factors IX and VIIa), interferon, and hormone-based cytokines. The goal of the current application is to develop a technology that can improve the production volume and pharmacy price of these difficult to manufacture recombinant biologics. Expression Therapeutics has developed a new baby hamster kidney-derived (BHK) cell line, designated BHK-MS, which can be cultured and expanded indefinitely in serum and blood product-free medium under suspension bioreactor conditions. In the current studies, we are 1) developing and validating the suspension adapted BHK-MS cell line for superior coagulation factor VIII and factor IX production, and 2) performing a pilot-scale manufacturing run and analyzing the biochemical and pharmaceutical characteristics of BHK-MS biosynthesized recombinant factor VIII and factor IX, which will serve as a model, difficult to manufacture biopharmaceuticals. PUBLIC HEALTH RELEVANCE: The long-term commercial objective of the current project is to commercialize a suspension cell-based biomanufacturing platform designed for improved volumetric productivity of complex, hard to manufacture biologics. Low-yield manufacturing processes currently in place often result in limited product availability and high-product costs. Expression Therapeutics has developed a patent-pending cell line designed for efficient manufacturing of recombinant biologics. The studies proposed herein are designed to provide proof-of-concept of the efficiency and scalability, as well as, rigorous preclinical validation and characterization of this novel manufacturing platform.